In vitro reconstructed models of human tissues are physiologically relevant alternatives to animal tests in many areas. They have been shown to reliably predict toxicity, interaction with pathogens, absorption and metabolism of pharmaceuticals and other substances as well as other processes. The prediction of acute respiratory toxicity (ART) and irritation is currently largely dependent on the use of animal models, which is problematic on both physiological and ethical grounds. The goal of this work was to develop physiologically relevant ART in vitro tests utilizing the EpiAirway? tracheobronchial tissue model, to demonstrate correlation to OECD-accepted GHS categorization. Test articles (n=53) were applied to tissues produced at two separate facilities with two ART protocols, the Direct Application Protocol (DAP) for exposure to mists/sprays, and the Vapor Cap Protocol (VCP) for exposure to vapors/volatile liquids. The effects on tissue viability (MTT assay) and barrier properties (TEER) were determined. The effective doses were interpolated for the DAP and VCP methods and correlated to the GHS categories. Using MTT, the Sensitivity/Specificity/Accuracy (S/S/A) of DAP method were 63.5/76.1/69.8% (lab1) and 63.8/76.1/70.0% (lab2) - R² =0.91. The VCP showed S/S/A of 70.8/83.2/77.0 (lab1) and 71.9/83.2/77.5% (lab2) - R² =0.93. Using the MTT assay, both VCP and DAP demonstrated good predictivity of GHS categories and high interlaboratory reproducibility. Both protocols provide robust, efficient, physiologically relevant, organ-specific in vitro tests that can improve the predictivity of human responses and reduce the number of animals used to assess respiratory toxicity.